Veksler V I, Kuznetsov A V, Sharov V G, Kapelko V I, Saks V A
Biochim Biophys Acta. 1987 Jun 29;892(2):191-6. doi: 10.1016/0005-2728(87)90174-5.
Respiratory parameters of cardiac mitochondria were determined in the bundles of cardiac fibers skinned by using saponin that specifically removed sarcolemma, but left intracellular structures intact. In the assay medium which simulated the ion composition of cardiac cytoplasm maximal value of state 3 oxygen consumption per mol cytochromes aa3 was close to that value for isolated mitochondria. Ischemia and isopreterenol treatment were found to affect respiratory parameters of mitochondria in saponin-skinned fibers, among them creatine-stimulated respiration decreased most significantly, (3-4)-times under these conditions. The method described can be easily applied for determination of the mitochondrial respiratory parameters in small (5-10 mg) biopsy samples from human heart.
通过使用皂角苷处理心肌纤维束来测定心肌线粒体的呼吸参数。皂角苷能特异性去除肌膜,但使细胞内结构保持完整。在模拟心肌细胞质离子组成的测定培养基中,每摩尔细胞色素aa3的状态3氧消耗最大值接近分离线粒体的该值。发现缺血和异丙肾上腺素处理会影响皂角苷处理的纤维中线粒体的呼吸参数,其中肌酸刺激的呼吸在这些条件下下降最为显著,降低了3 - 4倍。所描述的方法可轻松应用于测定来自人心脏的小(5 - 10毫克)活检样本中的线粒体呼吸参数。